CN111334403A - Micro-bubble generation chip based on micro-fluidic and preparation method and application thereof - Google Patents
Micro-bubble generation chip based on micro-fluidic and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a micro-bubble generation chip based on micro-fluidic and a preparation method and application thereof, relating to the technical field of micro-fluidic. The micro-fluidic-based micro-bubble generation chip comprises a substrate and a micro-fluidic control cavity layer arranged opposite to the substrate, wherein the micro-fluidic control cavity layer is provided with a micro-fluidic cavity, a microporous structure layer is arranged between the substrate and the micro-fluidic cavity layer, and the microporous structure layer is provided with a plurality of micropores; the microporous structure layer is seamlessly combined with the substrate, the microfluidic cavity layer is bonded with the microporous structure layer, and the microfluidic cavity corresponds to a plurality of micropores. The microbubble generation chip has a simple structure, forms a large number of hemispherical microbubbles on the premise of not influencing the physical and chemical properties of liquid in a microfluidic cavity channel, widens the application of the microbubble generation chip, can be used for enriching or screening the same or different particles, and can also be used for researching a microporous sonoluminescence mechanism.
Description
Technical Field
The invention relates to the technical field of microfluidics, in particular to a microbubble generation chip based on microfluidics and a preparation method and application thereof.
Background
With the increasing maturity of micro-nano processing technology, micro-fluidic chips (also called Lab-on-a-chips) are rapidly developed, which are a kind of micro reaction or analysis system using micro-channel network as structural feature, and have the advantages of fast analysis speed, less reagent consumption, low use cost, easy integration and automation, etc., so the micro-fluidic chips have been widely applied in the research of the fields of chemistry, biology, medicine, etc.
In the microfluidic channel, the formation method of the microbubbles is various, and the formation modes of external injection (Marmottant P, et al. nature,2003,423(6936): 153-. The sizes of the microbubbles are difficult to control by injecting air into the outside to form the microbubbles, the microbubbles formed by laser induction are generated by instantly generating a large amount of thermal cavitation, the microstructures are microstructures with notches arranged on the side surfaces of the microfluidic channels, and the formed microbubbles are semi-cylindrical microbubbles on the side surfaces.
The microbubbles formed by these prior structures and methods have certain limitations that are not conducive to further applications.
Therefore, it is desirable to obtain a new mode of microbubble formation that can solve at least one of the above-described problems.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
One of the purposes of the invention is to provide a micro-fluidic-based microbubble generation chip which is simple in structure, can form a large number of hemispherical microbubbles on the premise of not influencing the physicochemical properties of liquid in a micro-fluidic cavity channel, is convenient to apply, can be used for enrichment or screening of the same or different particles, and can also be used for researching a sonoluminescence mechanism.
The second objective of the present invention is to provide a method for preparing the micro-fluidic micro-bubble generating chip, which has the same advantages as the micro-fluidic micro-bubble generating chip, and has the advantages of simple preparation method and low cost.
The invention also aims to provide the application of the micro-fluidic micro-bubble generating chip or the micro-bubble generating chip prepared by the preparation method of the micro-fluidic micro-bubble generating chip in micro-fluidic mixing or enrichment screening of cells, microspheres and microorganisms.
The fourth purpose of the present invention is to provide an application of the micro-bubble generation chip based on the micro-fluidic system or the micro-bubble generation chip prepared by the preparation method of the micro-bubble generation chip based on the micro-fluidic system in multi-micro-bubble sonoluminescence.
In order to achieve the above purpose of the present invention, the following technical solutions are adopted:
in a first aspect, a micro-fluidic-based micro-bubble generation chip is provided, which comprises a substrate and a micro-fluidic control channel layer arranged opposite to the substrate, wherein the micro-fluidic control channel layer is provided with a micro-fluidic channel, a microporous structure layer is arranged between the substrate and the micro-fluidic channel layer, and the microporous structure layer is provided with a plurality of micropores; the microporous structure layer is seamlessly combined with the substrate, the microfluidic cavity layer is bonded with the microporous structure layer, and the microfluidic cavity corresponds to a plurality of micropores.
Preferably, on the basis of the technical scheme provided by the invention, the micropores of the micropore structure layer are arranged in an array;
preferably, the diameter of the micropores arranged in the array is the same or varies in gradient along the same direction.
Preferably, on the basis of the technical scheme provided by the invention, two ends of the microfluidic channel layer are respectively and independently provided with a sample port;
preferably, a sample dispersing structure is further arranged in the microfluidic channel of the microfluidic channel layer.
Preferably, on the basis of the technical scheme provided by the invention, the microfluidic channel layer and the microporous structure layer are respectively and independently provided with a positioning structure.
Preferably, on the basis of the technical scheme provided by the invention, the material of the micro-fluidic based micro-bubble generation chip comprises one of a silicon material, a glass quartz material, an organic high polymer material or a paper material;
preferably, the substrate is made of a glass quartz material;
preferably, the materials of the microfluidic channel layer and the microporous structure layer are both independently organic high polymer materials, preferably siloxane polymer materials, and further preferably PDMS materials.
In a second aspect, a method for preparing the microfluidic-based microbubble generation chip is provided, which comprises the following steps:
independently preparing a micro-fluidic control cavity layer with a micro-fluidic cavity channel and a microporous structure layer with a plurality of micropores, bonding the micro-fluidic cavity layer and the microporous structure layer to enable the micro-fluidic cavity channel to correspond to the positions of the micropores, and seamlessly bonding the microporous structure layer on a substrate to obtain the micro-fluidic-based microbubble generation chip.
Preferably, on the basis of the technical scheme provided by the invention, the processing methods of the micro-fluidic channel layer and the micro-porous structure layer independently comprise a photolithography method, a laser etching method, a template casting method or a template hot pressing method, and preferably the photolithography method.
Preferably, on the basis of the technical scheme provided by the invention, the preparation method of the micro-fluidic-based microbubble generation chip comprises the following steps:
(a) independently preparing a micro-fluidic channel layer and a microporous structure layer: spin-coating a photoresist on a substrate, and obtaining a required photoresist structure on the substrate by utilizing a photoetching process; then mixing PDMS with a hardening agent, pouring the mixture onto a substrate with a photoresist structure, and curing to obtain a micro-fluidic cavity layer and a microporous structure layer respectively;
(b) punching holes at two ends of a micro-fluidic channel of the micro-fluidic channel layer;
(c) and independently carrying out oxygen plasma treatment on the micro-flow control cavity layer and the microporous structure layer, bonding the micro-flow control cavity layer and the microporous structure layer together, and seamlessly bonding the micro-flow control cavity layer and the microporous structure layer on the substrate to obtain the micro-flow control based microbubble generation chip.
In a third aspect, the micro-fluidic micro-bubble generating chip or the micro-bubble generating chip prepared by the preparation method of the micro-fluidic micro-bubble generating chip is applied to micro-fluidic mixing or enrichment screening of cells, microspheres and microorganisms.
In a fourth aspect, an application of the micro-bubble generation chip based on the micro-fluidic system or the micro-bubble generation chip prepared by the preparation method of the micro-bubble generation chip based on the micro-fluidic system in multi-micro-bubble sonoluminescence is provided.
Compared with the prior art, the invention has the following beneficial effects:
the micro-fluidic micro-bubble generation chip has a simple structure and low cost, and the micro-pore structure layer with a plurality of micro-pores is arranged below the micro-fluidic cavity channel, so that after the chip is filled with liquid, the liquid flows through the micro-pore structure to form a liquid-air film due to the existence of the surface tension of the liquid, a micro-bubble is generated on each micro-pore, a plurality of micro-pores generate a plurality of micro-bubbles, and the chip with the structure can form a large number of hemispherical micro-bubbles on the premise of not influencing the physical and chemical properties of the liquid in the micro-fluidic cavity channel. The chip utilizing the structure of the invention can expand the application limitation of the traditional structure, and has wider application prospect. Under the external stimulation, the multi-microbubble generates the co-vibration, so that not only can the mixing of microfluid be realized, but also the microbubble vibration amplitude can be changed by adjusting the input energy or frequency, different particles are captured, and the enrichment and the screening of cells, microspheres or microorganisms are realized. The microbubble is violently and periodically contracted under the excitation of an external signal, so that picosecond-level flash can be generated, the microbubble sonoluminescence can be realized, and the research on the theoretical mechanism and the application of the microbubble sonoluminescence is promoted. In addition, the chip with the structure can enable the multi-microbubble to generate resonance under lower input energy (below 15W of input power), and effectively avoids the heat effect of the chip.
Drawings
FIG. 1 is a schematic structural diagram of a microbubble generation chip according to an embodiment of the present invention in an isolated state;
FIG. 2 is a schematic diagram of a backside structure of the micro-fluidic channel layer of FIG. 1;
FIG. 3 is a plan view of a microporous structure layer according to one embodiment of the present invention;
FIG. 4 is a plan view of a microfluidic channel layer according to one embodiment of the present invention;
FIG. 5 is a plan view of a microbubble generation chip according to an embodiment of the present invention;
FIG. 6 is an enlarged schematic view of the pore arrangement of the microporous structure layer according to one embodiment of the present invention;
FIG. 7 is an enlarged schematic view of the pore arrangement of the microporous structure layer according to another embodiment of the present invention;
FIG. 8 is a schematic view of a sample distribution structure according to an embodiment of the present invention;
FIG. 9 is a schematic view of a microporous structure layer and a microfluidic channel layer according to an embodiment of the present invention;
FIG. 10 is a schematic diagram showing the capture of 1 μm-diameter PS beads on the microbubble generation chip of example 2;
FIG. 11 is a diagram showing the capture of PS beads with a diameter of 10 μm on the microbubble generation chip of example 2;
FIG. 12 shows the capture of PS beads of different diameters at different positions.
The figure is as follows: 100-a substrate; 200-a microporous structural layer; 210-microwells; 300-microfluidic channel layer; 310-microfluidic channels; 320-sample port; 330-sample distribution structure; 340-positioning structure.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to examples, but it will be understood by those skilled in the art that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
According to one aspect of the invention, a micro-fluidic-based micro-bubble generation chip is provided, which comprises a substrate and a micro-fluidic control cavity channel layer arranged opposite to the substrate, wherein the micro-fluidic control cavity channel layer is provided with a micro-fluidic control cavity channel; the microporous structure layer is seamlessly combined with the substrate, the microfluidic cavity layer is bonded with the microporous structure layer, and the microfluidic cavity corresponds to the positions of the micropores.
As shown in fig. 1 to 5, the microbubble generation chip sequentially includes a substrate 100, a microporous structure layer 200, and a microfluidic channel layer 300 from bottom to top.
The materials of the substrate 100, the microporous structure layer 200, and the micro-fluidic channel layer 300 independently include, but are not limited to, silicon material, glass quartz material, organic polymer material, or paper material.
The material of the substrate 100 is exemplified by, for example, a glass quartz material, and an exemplary substrate is a glass slide.
The microporous structure layer 200 is a sheet having a plurality of micropores 210, and the arrangement and size of the micropores are not limited, and the sizes of the micropores may be the same or different.
The preparation method of the microporous structure layer is not limited, and can be carried out by adopting a conventional processing mode in the field of microfluidic chips.
The material of the microporous structure layer is not limited, and the material of the microporous structure layer is exemplified by, for example, an organic high polymer material, which may be a siloxane polymer material, and an exemplary material of the microporous structure layer is PDMS (polydimethylsiloxane).
The microfluidic channel layer 300 is a sheet having a microfluidic channel 310 structure, and the shape and size of the microfluidic channel are not limited, and the microfluidic channel is used for passing liquid, such as water, PBS buffer solution, blood or other liquid to be detected.
The preparation method of the micro-fluidic channel layer is not limited and can be carried out by adopting a conventional processing mode in the field of micro-fluidic chips.
The material of the micro-fluidic channel layer is not limited, and the material of the micro-fluidic channel layer is, for example, an organic polymer material, and an exemplary material of the micro-fluidic channel layer is PDMS (polydimethylsiloxane).
The microfluidic cavity layer is bonded with the microporous structure layer, the bonding method is not limited, and the bonding method can be performed by adopting a conventional bonding method in the field of microfluidic chips, for example, a thermal bonding mode, an anodic bonding mode or a low-temperature bonding mode can be adopted, the microfluidic cavity is corresponding to the positions of the micropores, the plurality of microporous structures are aligned to correspond to the microfluidic cavity structure during bonding, namely the plurality of micropores fall into the range of the microfluidic cavity, and liquid can cover the micropores when flowing through the cavity.
The microporous structure layer is seamlessly combined with the substrate, wherein the seamless combination means that the microporous structure layer is completely attached to the substrate, no gap or air exists between the two layers, and finally the microbubble generating chip is formed.
In one embodiment, as shown in fig. 4, the microfluidic channel layer has sample ports 320 at both ends of the microfluidic channel independently.
The micro-bubble generating chip based on micro-flow control of the invention is provided with the microporous structure layer with a plurality of micropores below the micro-flow control cavity channel, so that after the chip is filled with liquid, the liquid can form a liquid-air film after flowing through the microporous structure due to the existence of the surface tension of the liquid, thereby generating a micro-bubble on each micropore, and the plurality of micropores generate a plurality of micro-bubbles. The micro-bubble generating device has the advantages that stable cavitation is generated in the micro-bubble through the pressure difference between air surrounded by liquid and surrounding liquid through external stimulation (such as ultrasound), resonance is generated, and a micro-flow field is generated. In addition, the multi-microbubble is severely and periodically contracted under the excitation of an external signal, so that picosecond-level flash can be generated, the multi-microbubble sonoluminescence can be realized, and the research on the theoretical mechanism and the application of the multi-microbubble is promoted.
The microbubble generation chip has the advantages of simple structure, low cost and low input energy, and the input power is generally below 15W, so that the microbubbles can generate resonance, and the heat effect of the chip is effectively avoided.
In one embodiment, the micropores of the microporous structure layer are arranged in an array.
The array arrangement means that the arrangement mode of the microbubbles is regular. The microbubbles may be the same size or different sizes.
The arrangement in an array configuration can achieve high-throughput, large-scale capture of particles, and especially a graded array configuration can achieve capture of different particles at different locations.
In an exemplary embodiment, as shown in FIG. 6, the microwells are arranged in an array having the same diameter, i.e., an array of microbubbles having the same diameter.
The exemplary microwell array has two adjacent rows of microwells in a staggered arrangement.
In another exemplary embodiment, as shown in fig. 7, the diameters of the micropores arranged in the array vary in a gradient manner along the same direction, where the same direction may be the length direction (X direction) of the plane of the microporous structure layer, or the width direction (Y direction) of the plane of the microporous structure layer, and the gradient variation may increase in a gradient manner or decrease in a gradient manner, that is, the diameter of the microbubble array becomes gradually larger or smaller.
Because the micro bubbles with different diameters have different vibration amplitudes, when the micropore array is a micropore with the same diameter, the same particles can be captured, and when the micropore array is a micropore with the gradually changed diameter, the functions of capturing, screening and the like of different particles can be realized.
In one embodiment, as shown in fig. 8, the microfluidic channel of the microfluidic channel layer 300 further has a sample distribution structure 330.
The sample dispersing structure is integrally formed when the microfluidic cavity layer is prepared, the sample dispersing structure is arranged in the cavity close to the sample port, the shape of the sample dispersing structure is not limited, and only the liquid enters the cavity from the sample port and then has a shunting effect.
By arranging the sample dispersing structure, substances (such as cells, microorganisms and the like) in the liquid are uniformly distributed in the microfluidic cavity.
In one embodiment, the positioning structure 340 is disposed on each of the micro-fluidic channel layer 300 and the microporous structure layer 200.
Because the cavity and the micropore structure can be seen clearly under a microscope, in order to realize quick and accurate bonding, positioning structures are independently arranged on the micro-fluidic cavity layer and the micropore structure layer, and accurate bonding can be realized by directly aligning the positioning structures, so that the positions of a plurality of micropores correspond to the micro-fluidic cavity structure, and the plurality of micropores fall into the micro-fluidic cavity range.
An exemplary micro-fluidic-based micro-bubble generation chip sequentially comprises a substrate, a micro-porous structure layer and a micro-fluidic cavity layer from bottom to top, wherein the micro-porous structure layer is provided with a plurality of micro-pores which are arranged in an array mode, the diameters of the micro-pores are the same or change along the same direction in a gradient mode, the micro-fluidic cavity layer is provided with a micro-fluidic cavity channel, two ends of the micro-fluidic cavity channel are respectively and independently provided with a sample port, a sample dispersing structure is further arranged in the micro-fluidic cavity channel, the micro-fluidic cavity channel and the micro-porous structure layer are respectively and independently provided with a positioning structure, the micro-fluidic cavity channel of the micro-fluidic cavity layer is correspondingly bonded with the positions of the micro-.
When the micro-fluidic device is used, a sample to be detected is injected into a micro-fluidic cavity channel through a sample port by an injector through a hose, the sample to be detected flows through a micro-porous array after passing through a sample dispersion structure, an air-liquid film is formed at the position of the micro-porous structure due to the surface tension of liquid, other gases can be filled in advance, the air at the structure is evacuated, micro-bubbles generate co-vibration under the excitation of external ultrasound, so that a micro-fluidic field is generated, micro-flow can be used for controlling and screening organisms in the cavity channel without direct contact and damage, the magnitude of acting force can be controlled by adjusting the magnitude of an input signal, and the vibration amplitude of the micro-bubbles can be controlled, so that the enrichment and screening of cells, microspheres or microorganisms and the mixing of micro-fluid; due to the periodic rapid diastolic motion of the surface of the microbubble, the microbubble can burst picosecond-level flash light, and the microbubble can be used for researching the multi-microbubble sonoluminescence mechanism.
According to a second aspect of the present invention, there is provided a method for preparing a microfluidic-based microbubble generation chip, comprising the steps of: independently preparing a micro-fluidic control cavity layer with a micro-fluidic cavity channel and a microporous structure layer with a plurality of micropores, bonding the micro-fluidic cavity layer and the microporous structure layer to enable the micro-fluidic cavity channel to correspond to the positions of the micropores, and seamlessly bonding the microporous structure layer on a substrate to obtain the micro-fluidic-based microbubble generation chip.
Methods for fabricating the microfluidic channel layer and the microporous structure layer include, but are not limited to, photolithography, laser etching, template casting, or template hot pressing, such as photolithography. The micro-pore structure processed by the soft lithography technology is more convenient than other existing methods, and the size and the position of the micro-bubble can be designed according to needs.
The bonding method is not limited, and thermal bonding, anodic bonding, low-temperature bonding, or the like can be used.
The preparation method of the microbubble generating chip is simple, low in cost and effective.
FIG. 9 is a schematic diagram of one embodiment of a microporous structure layer and a microfluidic channel layer, as shown in FIG. 9, an exemplary method comprising: and spin-coating photoresist on the substrate, curing, exposing under a mask, developing to leave a designed pattern, pouring PDMS into the substrate with the pattern structure, curing, and then removing the bonding.
The exposure and development principle is as follows: in the area irradiated by ultraviolet light, the inside of the photoresist generates a crosslinking reaction and is the area irradiated by light; the photoresist does not generate cross-linking reaction inside, so that the curing degree of the illuminated area is far greater than that of the non-illuminated area, after the developing solution is soaked and cleaned, the illuminated area is reserved, and other areas are dissolved.
An exemplary substrate is a silicon wafer, for example.
In one embodiment, the method for preparing a microfluidic-based microbubble generation chip comprises the steps of:
(a) independently preparing a micro-fluidic channel layer and a microporous structure layer: spin-coating a photoresist on a substrate, and obtaining a required photoresist structure on the substrate by utilizing a photoetching process; then mixing PDMS with a hardening agent, pouring the mixture onto a substrate with a photoresist structure, and curing to obtain a micro-fluidic cavity layer and a microporous structure layer respectively;
(b) punching holes at two ends of a micro-fluidic channel of the micro-fluidic channel layer;
(c) and independently carrying out oxygen plasma treatment on the micro-flow control cavity layer and the microporous structure layer, bonding the micro-flow control cavity layer and the microporous structure layer together, and seamlessly bonding the micro-flow control cavity layer and the microporous structure layer on the substrate to obtain the micro-flow control based microbubble generation chip.
According to a third aspect of the present invention, there is provided an application of the above-mentioned micro-fluidic based micro-bubble generating chip or the micro-bubble generating chip prepared by the above-mentioned micro-fluidic based micro-bubble generating chip in micro-fluidic mixing or enrichment screening of cells, microspheres and microorganisms.
The liquid flows through the micro-pores to form multi-micro-bubbles, the micro-bubbles are hemispherical, the micro-bubbles vibrate to generate a micro-flow field under the excitation of an external piezoelectric transducer, the generated flow field is symmetrical, symmetrical vortexes can be used for mixing the liquid, and when the two liquids flow into the micro-fluidic channel, the two liquids are fully mixed under the resonance of the multi-micro-bubbles.
The micro-bubbles with different diameters have different vibration amplitudes, the micro-bubbles with different diameters can be generated by controlling the diameters of the micro-pores, and different particles (such as cells, microspheres, microorganisms and the like) can be captured by the vibration of the micro-bubbles, so that the enrichment and screening effects are achieved.
According to a fourth aspect of the present invention, there is provided an application of the above-mentioned micro-fluidic based micro-bubble generation chip or the micro-bubble generation chip prepared by the above-mentioned micro-fluidic based micro-bubble generation chip in multi-micro-bubble sonoluminescence.
The multi-microbubble formed by the chip generates periodic rapid diastolic motion under the action of external excitation, and the microbubble vibrates more violently under the condition of increasing energy, so that the microbubble can burst picosecond-level flash light and can be used for multi-microbubble sonoluminescence.
In order to further understand the present invention, the following will explain the method and effects of the present invention in detail with reference to specific examples and comparative examples. The following examples are merely illustrative of the present invention and should not be construed as limiting the scope of the invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The glass slide is a high-light-transmission medical glass slide.
Example 1
A micro-bubble generation chip based on micro-flow control sequentially comprises a glass slide, a micro-pore structure layer and a micro-flow control cavity layer from bottom to top, wherein the micro-pore structure layer is provided with a plurality of micro-pores which are arranged in an array mode, the diameters of the micro-pores are 40 micrometers, the micro-flow control cavity layer is provided with micro-flow control cavity channels, two ends of each micro-flow control cavity channel are respectively and independently provided with a sample port, a sample dispersing structure is further arranged in each micro-flow control cavity channel, the micro-flow control cavity channels and the micro-pore structure layer are respectively and independently provided with a positioning structure, the micro-flow control cavity channels of the micro-flow control cavity layer are correspondingly bonded with a plurality of.
A preparation method of a micro-bubble generation chip based on micro-fluidic comprises the following steps:
1. separately preparing a microfluidic channel layer and a microporous structure layer:
(1) baking the clean silicon wafer on a hot plate at 95 ℃ for 30 min;
(2) after cooling, the negative photoresist SU-83025 is spin-coated for 15s at 500rpm and 30s at 2000rpm on a spin coater for spin-coating;
(3) baking the spin-coated silicon wafer on a hot plate at 95 ℃ for 30 min;
(4) after cooling, a film containing a micro-fluidic channel structure or a micro-pore structure is arranged right above the photoresist area, and the photoresist is exposed through a photoetching machine;
(5) baking on a hot plate at 95 deg.C for 15 min;
(6) uniformly mixing the PDMS main agent and the hardening agent in a mass ratio of 10: 1;
(7) pouring the mixture into a silicon chip containing a micro-fluidic channel structure or a microporous structure, vacuumizing for 15min, and removing bubbles in PDMS;
(8) curing at 80 ℃ for 1 h;
(9) uncovering the cured PDMS to independently obtain a micro-fluidic channel PDMS layer and a microporous structure PDMS layer, and punching holes at a channel liquid inlet and an outlet of the micro-fluidic channel PDMS layer by using a puncher with the aperture of 0.75 mm;
2. bonding:
(10) independently processing the micro-fluidic channel PDMS layer and the microporous structure PDMS layer for 30s by oxygen plasma, then bonding the micro-fluidic channel PDMS layer and the microporous structure PDMS layer together (the positions of the channel and the micropores can be corresponding through a positioning structure), and attaching the micro-fluidic channel PDMS layer and the microporous structure PDMS layer on a glass slide;
(11) and (3) baking the micro-bubble generating chip in an oven at 80 ℃ for 30min to obtain the micro-bubble generating chip based on the micro-flow control.
Example 2
A micro-bubble generation chip based on micro-flow control sequentially comprises a glass slide, a micro-pore structure layer and a micro-flow control cavity layer from bottom to top, wherein a plurality of micro-pores are arrayed on the micro-pore structure layer, the diameters of the micro-pores are increased to 60 microns from minimum 20 microns along the length direction of the micro-pore structure layer in a group by taking three micro-pores as a group according to a gradient of 10 microns, a micro-flow control cavity channel is arranged on the micro-flow control cavity layer, sample ports are respectively and independently arranged at two ends of the micro-flow control cavity channel, a sample dispersing structure is further arranged in the micro-flow control cavity channel, positioning structures are respectively and independently arranged on the micro-flow control cavity channel layer and the micro-pore structure layer, the micro-flow control cavity channel of the micro-.
The preparation method of the micro-fluidic based micro-bubble generation chip is the same as that of example 1.
Example 3
A micro-bubble generation chip based on micro-flow control sequentially comprises a glass slide, a micro-pore structure layer and a micro-flow control cavity layer from bottom to top, wherein a plurality of micro-pores which are randomly distributed are arranged on the micro-pore structure layer, micro-flow control cavity channels are arranged on the micro-flow control cavity layer, sample ports are respectively and independently arranged at two ends of each micro-flow control cavity channel, a sample dispersing structure is further arranged in each micro-flow control cavity channel, positioning structures are respectively and independently arranged on the micro-flow control cavity channel layer and the micro-pore structure layer, the micro-flow control cavity channels of the micro-flow control cavity layer are correspondingly bonded with the positions of the micro-pores of the micro-pore.
The preparation method of the micro-fluidic based micro-bubble generation chip is the same as that of example 1.
Comparative example 1
This comparative example is different from example 1 in that the microporous structure layer has a single micropore having a diameter of 40 μm. The preparation method is correspondingly adjusted.
Application example PS bead Capture test
PS (Polystyrene, abbreviated as PS) bead trapping test was performed using the microbubble generation chips of example 2 and comparative example 1, respectively, as follows:
through the peristaltic pump, inject the PS bobble diluent into the chamber way, couple PZT and slide glass together with the ultrasonic couplant, signal generator input energy, the work of excitation PZT, the vibration of PZT arouses the vibration of microbubble, because the second order radiation force that the particle of different diameters received in the microbubble department of different diameters and the sound miniflow size are different, consequently can be so that the PS bobble of different diameters is caught in different positions to realize the screening.
The capture of PS beads with a diameter of 1 micron in the graded microbubble array structure of example 2 is shown in fig. 10, and as can be seen from fig. 10, PS beads are captured mainly at a microbubble diameter of 20 microns.
The capture of PS beads with a diameter of 10 microns in the graded microbubble array structure of example 2 is shown in fig. 11, and as can be seen from fig. 11, PS beads are captured mainly at a microbubble diameter of 50 microns.
FIG. 12 shows the capture of PS beads of different diameters at different positions.
As can be seen from fig. 12, particles of 1 μm and 10 μm are captured by microbubbles of different diameters due to the difference in diameter, and due to this difference, the device can be used to screen cells of different diameters for blood detection of disease.
While the single microbubble of comparative example 1 can only realize the screening of a single and small amount of particles, the PS globules with the diameter of 2 microns form a pair of symmetrical eddy currents in the acoustic microflow field generated by the microbubble of comparative example 1 with the diameter of 40 microns.
Therefore, the invention can realize high-throughput and large-scale screening.
While particular embodiments of the present invention have been illustrated and described, it would be obvious that various other changes and modifications can be made without departing from the spirit and scope of the invention. It is therefore intended to cover in the appended claims all such changes and modifications that are within the scope of this invention.
Claims (10)
1. A micro-fluidic-based micro-bubble generation chip is characterized by comprising a substrate and a micro-fluidic control cavity layer arranged opposite to the substrate, wherein the micro-fluidic control cavity layer is provided with a micro-fluidic cavity, a microporous structure layer is arranged between the substrate and the micro-fluidic control cavity layer, and the microporous structure layer is provided with a plurality of micropores; the microporous structure layer is seamlessly combined with the substrate, the microfluidic cavity layer is bonded with the microporous structure layer, and the microfluidic cavity corresponds to a plurality of micropores.
2. A microfluidic based microbubble generation chip according to claim 1, wherein the micropores of the microporous structure layer are arranged in an array;
preferably, the diameter of the micropores arranged in the array is the same or varies in gradient along the same direction.
3. The microfluidic based microbubble generation chip of claim 1, wherein the microfluidic channel layer has sample ports at both ends of the microfluidic channel independently;
preferably, a sample dispersing structure is further arranged in the microfluidic channel of the microfluidic channel layer.
4. The microfluidic based microbubble generation chip of any one of claims 1 to 3, wherein the microfluidic channel layer and the microporous structure layer are each independently provided with a positioning structure.
5. The microfluidic based microbubble generation chip according to any one of claims 1 to 3, wherein the substrate is made of a glass quartz material;
preferably, the materials of the microfluidic channel layer and the microporous structure layer are both independently organic high polymer materials, preferably siloxane polymer materials, and further preferably PDMS materials.
6. A method for preparing a microfluidic-based microbubble generation chip according to any one of claims 1 to 5, comprising the steps of:
independently preparing a micro-fluidic control cavity layer with a micro-fluidic cavity channel and a microporous structure layer with a plurality of micropores, bonding the micro-fluidic cavity layer and the microporous structure layer to enable the micro-fluidic cavity channel to correspond to the positions of the micropores, and seamlessly bonding the microporous structure layer on a substrate to obtain the micro-fluidic-based microbubble generation chip.
7. The method of claim 6, wherein the microfluidic channel layer and the microporous structure layer are processed independently by photolithography, laser etching, template casting, or template hot pressing, preferably photolithography.
8. The method for preparing a microfluidic-based microbubble generation chip according to claim 6, comprising the steps of:
(a) independently preparing a micro-fluidic channel layer and a microporous structure layer: spin-coating a photoresist on a substrate, and obtaining a required photoresist structure on the substrate by utilizing a photoetching process; then mixing PDMS with a hardening agent, pouring the mixture onto a substrate with a photoresist structure, and curing to obtain a micro-fluidic cavity layer and a microporous structure layer respectively;
(b) punching holes at two ends of a micro-fluidic channel of the micro-fluidic channel layer;
(c) and independently carrying out oxygen plasma treatment on the micro-flow control cavity layer and the microporous structure layer, bonding the micro-flow control cavity layer and the microporous structure layer together, and seamlessly bonding the micro-flow control cavity layer and the microporous structure layer on the substrate to obtain the micro-flow control based microbubble generation chip.
9. Use of a micro-fluidic based micro-bubble generating chip according to any one of claims 1 to 5 or a micro-bubble generating chip prepared by the method of preparing a micro-fluidic based micro-bubble generating chip according to any one of claims 6 to 8 in micro-fluidic mixing or enrichment screening of cells, microspheres and microorganisms.
10. Use of a microfluidic-based microbubble generation chip according to any one of claims 1 to 5 or a microbubble generation chip prepared by the method of preparing a microfluidic-based microbubble generation chip according to any one of claims 6 to 8 in microbubble sonoluminescence.
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